中文摘要：

目的：研究能量可控陡脉冲（energy controllable sleep pulse，ECSP）对细胞缝隙连接蛋白Cx43的影响。方法：处理组用不同剂量参数组合的ECSP对乳腺癌MDA—MB-231细胞处理5min后继续培养48h，对照组不予任何处理。2组均采用间接免疫荧光细胞化学法测定和激光共聚焦显做镜观察连接蛋白Cx43的定位。采用Western印迹法检测细胞内Cx43蛋白的含量，RT—PCR检测Cx43mRNA的表达。结果：对照组乳腺癌细胞间连接处有散在的明亮点状标记。ECSP处理2、3、4组经ECSP处理（条件分别为：电压100V，频率100Hz；电压100V，频率200Hz；电压150V，频率100Hz）后，继续培养的乳腺癌细胞间连接处标记斑点较对照组增多。Western印迹分析表明：ECSP处理2、3、4组Cx43蛋白条带亮度明显增加，与对照组相比差异有统计学意义（P〈0．05）。RT—PCR扩增产物中，ECSP处理2、3、4组的Cxa3基因条带亮度明显高于对照组，差异有统计学意义（P〈0．05）。ECSP处理1、5组（条件分别为：电压100V，频率50Hz；电压200V，频率200Hz）经ECSP处理后继续培养的乳腺癌细胞间连接处标记斑点与对照组无明显差异（P〉0．05）。ECSP处理1、5组Cx43蛋白条带与对照组相比，差异无统计学意义（P〉0．05）。RT—PCR扩增产物中，ECSP处理1、5组Cx43基因条带亮度与乳腺癌对照组比较，差异亦无统计学意义（P〉0．05）。结论：低能量可控陡脉冲可增强体外培养的乳腺癌细胞缝隙连接通讯功能，从而达到治疗恶性肿瘤的目的。

英文摘要：

Objective：To explore the effect of energy controllable steep pulse （ECSP） on gap junction intercellular communication （GJIC） protein. Methods：The breast carcinoma cells MDA-MB-231 in ECSP group were cultured for 48 h after treatment with different doses of ECSP for 5 min. The control group was cultured without any treatment. The localization of Cx43 proteins was determined by indirect immunofluorescent histochemical analysis and detected by laser confocal microscopy. The expression level of intracellular Cx43 protein was measured by Western blotting analysis and Cx43 mRNA level was tested by RT-PCR. Results：There were dispersed bright-labeled spots between breast carcinoma cellular junctions in the control group. ECSP treatment groups were given ECSP at different doses and frequencies （ voltage of 100 V and frequency of 100 Hz for group 2 ; voltage of 100 V and frequency of 200 Hz for group 3 ; voltage of 150 V and frequency of 100 Hz for group 4）. After treatment there were much more bright-labeled spots in the ECSP 2, 3, 4 groups than those in the control group. Western blotting analysis showed that the strips of Cx43 protein in ECSP 2, 3, 4 groups were brighter than that in the control group. There was a significant difference between ECSP 2, 3, 4 groups and control group （P 〈 0.05）. The strips of Cx43 gene in ECSP 2, 3,4 groups were brighter than that in control group in RT-PCR amplified products. There was a significant difference between ECSP 2, 3, 4 groups and control group （P 〈 0.05 ）. After treatment with ECSP at voltage of 100 V and frequency of 50 Hz （ ECSP 1 group） or at voltage of 200 V and frequency of 200 Hz （ ECSP 5 group） , there were no obvious differences between the ECSP 1 and 5 groups and control group in the labeled spots between intercellular junctions. There were no significant differences in the relative density of strips of Cx43 protein and Cx43 gene in RT-PCR amplified products between the ECSP 1 and 5 groups and the control group （ P 〉 0. 05 ?